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Effect of dexamethasone on development of in vitro-produced bovine embryos

机译:地塞米松对体外培养牛胚胎发育的影响

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Studies in somatic cells have shown that glucocorticoids such as dexamethasone (DEX) may trigger or prevent apoptosis depending on the cell type in culture. Because the dysregulation of apoptosis may lower in vitro embryo production efficiency, we sought to investigate the effects of supplementing IVC medium with DEX (0.1 mu g/mL) on embryo morphology, development kinetics, and apoptosis rates of in vitro-produced bovine preimplantation embryos. Embryo morphology was graded on Day 7, and development rates were assessed on Days 4 and 7 of IVC. Apoptosis was evaluated via annexin/propidium iodide staining under fluorescence microscopy where a cell labeled with annexin, propidium iodide, or both would be considered apoptotic. An embryo was counted in the apoptosis rates, if it displayed at least one such labeled cell. Although DEX supplementation did not reduce apoptosis rates, it had a positive impact on developmental kinetics and cell number both on Days 4 and 7 of embryo culture. Presumably, such effect resulted from increased cell proliferation rather than a direct inhibition of apoptosis. Further studies may evaluate the mechanisms by which glucocorticoids may affect embryo development, as DEX supplementation could become a tool to improve in vitro embryo yield in mammalian species.
机译:在体细胞中的研究表明,糖皮质激素如地塞米松(DEX)可能会触发或阻止细胞凋亡,具体取决于培养中的细胞类型。由于细胞凋亡的失调可能会降低体外胚胎的生产效率,因此,我们寻求研究在DEC(0.1μg / mL)中补充IVC培养基对体外培养的牛植入前胚胎的胚胎形态,发育动力学和细胞凋亡率的影响。 。在IVC的第7天对胚胎形态进行分级,并在第4天和第7天评估发育速度。在荧光显微镜下通过膜联蛋白/碘化丙啶染色评估凋亡,其中被膜联蛋白,碘化丙啶或两者标记的细胞被认为是凋亡的。如果胚胎显示出至少一个这样的标记细胞,则将其计数在凋亡率中。尽管补充DEX并不能降低细胞凋亡率,但在胚胎培养的第4天和第7天,它对发育动力学和细胞数量均具有积极影响。据推测,这种作用是由于细胞增殖增加而不是直接抑制细胞凋亡引起的。进一步的研究可能会评估糖皮质激素可能影响胚胎发育的机制,因为补充DEX可能会成为提高哺乳动物物种体外胚胎产量的工具。

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